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Targeted mutagenesis by homologous recombination in D. melanogaster

By Yikang S. Rong, Simon W. Titen, Heng B. Xie, Mary M. Golic, Michael Bastiani, Pradip Bandyopadhyay, Baldomero M. Olivera, Michael Brodsky, Gerald M. Rubin and Kent G. Golic

Abstract

We used a recently developed method to produce mutant alleles of five endogenous Drosophila genes, including the homolog of the p53 tumor suppressor. Transgenic expression of the FLP site-specific recombinase and the I-SceI endonuclease generates extrachromosomal linear DNA molecules in vivo. These molecules undergo homologous recombination with the corresponding chromosomal locus to generate targeted alterations of the host genome. The results address several questions about the general utility of this technique. We show that genes not near telomeres can be efficiently targeted; that no knowledge of the mutant phenotype is needed for targeting; and that insertional mutations and allelic substitutions can be easily produced

Topics: Research Paper
Publisher: Cold Spring Harbor Laboratory Press
Year: 2002
DOI identifier: 10.1101/gad.986602
OAI identifier: oai:pubmedcentral.nih.gov:186348
Provided by: PubMed Central
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